TP4 : Les protéines, expression primaire de l’information génétique

Constat initial :

  • Le phénotype d’un individu dépend de la fonction de la protéine synthétisée.
  • Une protéine est une macromolécule constituée d’acides aminés.
  • La synthèse d’une protéine dépend de l’expression d’un gène.
  • Un gène est un fragment d’ADN donc il est constitué d’une séquence de nucléotides.

Expérience 1 : Localisation de la synthèse des protéines

On veut connaître le lieu de synthèse des protéines à l’intérieur de la cellule, pour cela on réalise une série d’expériences sur l’acétabulaire, algue verte constituée d’une seule cellule et dont la forme générale des différentes espèces rappelle celle d’une ombrelle. Elle présente une tige élancée de 3 à 5 cm de long fixée sur les rochers par un rhizoïde (sortes de racines), au niveau duquel est contenu le noyau. A l’extrémité de la tige se forme une sorte de chapeau qui peut atteindre 1 cm de diamètre et dont la forme diffère d’une espèce à l’autre. Certaines portions de la cellule peuvent, après isolement, régénérer la cellule entière.

Protocole 1 : Des acétabulaires sont mises en culture dans un milieu contenant un acide aminé radioactif, la méthionine. Après 30 minutes on pratique une autoradiographie de ces cellules. Le résultat obtenu est schématisé ci-dessus. On vérifie après extraction et analyse chimique que la radioactivité est concentrée au niveau des protéines de l’acétabulaire.
Protocole 2 : Des acétabulaires sont mises en culture dans un milieu contenant de la thymine radioactive. Après 30 minutes on pratique une autoradiographie de ces cellules. Le résultat obtenu est schématisé ci-dessus. On vérifie après extraction et analyse chimique que la radioactivité est concentrée au niveau des molécules d’ADN de l’acétabulaire.

[important]

Q1 : Déduire de l’analyse de l’expérience ci-dessus, la localisation de la synthèse des protéines dans la cellule.

Partie commune Q2 : Quel problème biologique découle des informations apportées par ces documents sur la synthèse des protéines ?

Partie commune Q3 : Énoncer une hypothèse et ses conséquences vérifiables permettant de répondre à ce problème.

Q4 : L’ensemble des expériences suivantes vont permettre d’éprouver l’hypothèse et ainsi de comprendre le mécanisme de la synthèse des protéines. Faire l’analyse des ces expériences et conclure en expliquant ce mécanisme.

[/important]

Expérience 2 : Observation microscopique de cellules colorées au vert de méthyle-pyronine

Protocole :

  1. Prélever un fragment de l’épiderme interne d’une écaille d’oignon blanc et la placer dans les colorants pendant 2 minutes.
  2. Rincer ensuite l’échantillon dans le deuxième verre de montre.
  3. Monter l’échantillon rincé entre lame et lamelle dans une goutte d’eau et observer au microscope.
  4. Prendre une photo de l’observation.
  5. Dans un document OpenOffice ou Word, légender la photographie.
  6. Interpréter les résultats de la coloration.

Des analyses fines des constituants cellulaires montrent que les cellules contiennent divers types d’acides nucléiques dont l’ADN. Le vert de méthyle-pyronine est un colorant spécifique des acides nucléiques. Le vert de méthyle colore l’ADN en bleu vert et la pyronine colore une autre famille d’acides nucléiques, les ARN ou Acides Ribonucléiques, en rose.

Expérience 3 : Expérience historique sur les acétabulaires

Une acétabulaire en cours de croissance est découpée en deux fragments. Le fragment nucléé régénère le chapeau. Le fragment énucléé régénère un chapeau  de petite taille puis meurt. La régénération du chapeau nécessite la synthèse de protéines.

On  a mis en évidence dans le cytoplasme des cellules, un acide nucléique : l’ARN (acide ribonucléique). La même expérience que précédemment est réalisée mais chaque fragment est traité par l’ARNase, enzyme qui détruit  spécifiquement les molécules d’ARN. Aucun des deux  fragments ne régénère  un  chapeau.

Si on arrête l’action de l’ARNase, seul le fragment nucléé régénère un chapeau.

Expérience 4 : Étude du document 2 p.54

Expérience 5 : Comparer l’ARN et l’ADN

Expérience 5 : Comprendre la synthèse des protéines

[warning]

Exercice complémentaire :

[/warning]

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